| pEgr-sHemopexin重组质粒的构建及体外辐射诱导表达 |
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| 引用本文： | 王贵全,许传杰,杨文,朴春姬,董震.pEgr-sHemopexin重组质粒的构建及体外辐射诱导表达[J].吉林大学学报(医学版),2005,31(2):236-238. |
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| 作者姓名： | 王贵全 许传杰 杨文 朴春姬 董震 |
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| 作者单位： | 吉林大学中日联谊医院耳鼻咽喉-头颈外科,吉林,长春,130031;吉林大学第二医院信息中心,吉林,长春,130041;吉林大学第二医院信息中心,吉林,长春,130041;吉林大学公共卫生学院,卫生部放射生物学重点实验室,吉林,长春,130021;吉林大学中日联谊医院耳鼻咽喉-头颈外科,吉林,长春,130031 |
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| 基金项目： | 吉林省科技厅社会发展基金 |
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| 摘 要： | 目的：克隆小鼠分泌型Hemopexin (sPEX) 编码区的cDNA序列，构建含Egr-1启动子的pEgr-sPEX真核表达载体，检测辐射诱导重组质粒在B16F10细胞中的表达。
方法：用RT-PCR从NIH3T3细胞中扩增出sPEX，经测序证实后，构建pEgr-sPEX重组质粒，以脂质体转染B16F10细胞，用Western blotting方法检测B16F10细胞上清中辐射诱导PEX的表达。
结果：测序表明扩增的sPEX cDNA序列与GenBank中登录的序列完全一致，sPEX cDNA正确插入表达载体pcDNA3.1-Egr-1，转染入B16F10细胞内PEX基因在体外辐射诱导下成功获得表达。结论：成功地克隆了小鼠sPEX基因，并在体外证实了pEgr-sPEX具有辐射诱导表达特性。
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| 关 键 词： | Hemopexin操纵子 黑色素瘤 基因表达 质粒 |
| 文章编号： | 1671-587X(2005)02-0236-03 |
| 收稿时间： | 2004-10-19 |
| 修稿时间： | 2004年10月19日 |
Construction and expression of pEgr-sHemopexin recombinant plasmid induced by ionizing radiation in vitro |
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| WANG Gui-quan,XU Chuan-jie,YANG Wen,PIAO Chun-ji,DONG Zhen.Construction and expression of pEgr-sHemopexin recombinant plasmid induced by ionizing radiation in vitro[J].Journal of Jilin University: Med Ed,2005,31(2):236-238. |
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| Authors: | WANG Gui-quan XU Chuan-jie YANG Wen PIAO Chun-ji DONG Zhen |
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| Institution: | 1.Department of Otolaryngology and Head-Neck Surgery, China-Japan Union Hospital, Jilin University,Changchun 130031,China；2. Department of Information Center, Second Hospital, Jilin University,Changchun 130041,China;3. MH Radiobiology Research Unit,School of Public Health, Jilin University, Changchun 130021,China |
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| Abstract: | Objective To clone mouse secretable Hemopexin(sPEX) cDNA, construct pEgr-sPEX recombinant plasmid and detect the expression of recombinant plasmid in B16F10 cells. Methods Hemopexin cDNA was amplified from the NIH3T3 cells by RT-PCR. After the cDNA identified by sequencing, the pEgr-sPEX recombinant plasmid was constructed and the plasmid was transfected into B16F10 cells with liposome and the expression of PEX induced by ionizing radiation in B16F10 cells was detected by Western blotting. Results The sequencing results proved the cloned sPEX cDNA to be completely identical with that reported in the GenBank. The mouse sPEX cDNA was inserted correctly into expression vector and expressed successfully. Conclusion The mouse sPEX cDNA is cloned successfully and it is confirmed that pEgr-sPEX possesses the radiation inducing expression characteristics in vitro. |
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| Keywords: | Hemopexin operon melanoma gene expression plasmids |
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